Identification of hemiclonal reproduction in three species of Hexagrammos marine reef fishes

Natural hybrids between the boreal species Hexagrammos octogrammus and two temperate species Hexagrammos agrammus and Hexagrammos otakii were observed frequently in southern Hokkaido, Japan. Previous studies revealed that H. octogrammus is a maternal ancestor of both hybrids; the hybrids are all fertile females and they frequently breed with paternal species. Although such rampant hybridization occurs, species boundaries have been maintained in the hybrid zone. Possible explanations for the absence of introgressions, despite the frequent backcrossing, might include clonal reproduction: parthenogenesis, gynogenesis and hybridogenesis. The natural hybrids produced haploid eggs that contained only the H. octogrammus genome (maternal ancestor) with discarded paternal genome and generated F₁‐hybrid type offspring by fertilization with the haploid sperm of H. agrammus or H. otakii (paternal ancestor). This reproductive mode was found in an artificial backcross hybrid between the natural hybrid and a male of the paternal ancestor. These findings indicate that the natural hybrids adopt hybridogenesis with high possibility and produce successive generations through hybridogenesis by backcrossing with the paternal ancestor. These hybrids of Hexagrammos represent the first hybridogenetic system found from marine fishes that widely inhabit the North Pacific Ocean. In contrast with other hybridogenetic systems, these Hexagrammos hybrids coexist with all three ancestral species in the hybrid zone. The coexistence mechanism is also discussed.     

Microsatellite markers and multiple paternity in a paternal care fish, Hexagrammos otakii

The fertilization success of sneaking behavior in a paternal care fish, Hexagrammos otakii, was estimated using microsatellites as genetic markers. Sneaking behavior was observed using a video camera placed near the nest of a breeding territorial male. Two egg masses fertilized by different patterns of sperm emission by the territorial male and sneaker males were examined for paternity discrimination. When a sneaker emitted a large amount of sperm after sperm emission by the territorial male, the sneaker fertilized more eggs than the territorial male. When the territorial male and a sneaker alternately emitted semen, the territorial male fertilized the most eggs. These results suggest that both the amount of emitted sperm and the timing of sperm emission are important factors in sperm competition. Received: November 18, 1999 / Accepted: January 17, 2000     

Mating systems of two midwater-spawning cichlids,Cyprichromis microlepidotus andParacyprichromis brieni,in Lake Tanganyika

The breeding habits of 2 maternal mouthbrooding cichlids,Cyprichromis microlepidotus andParacyprichromis brieni, were investigated in Lake Tanganyika. Although spawning on the substrate in the male's mating territory is prevalent in maternal mouthbrooders, bothC. microlepidotus andP. brieni spawned in the water column. MaleC. microlepidotus established their mating territories in the open water column, while maleP. brieni did so around fixed spawning sites near a vertical rock surface. In both species, females visited male mating territories, departing soon after spawning and collecting the eggs. Sneaking, which was observed only inP. brieni, may be attributed to the presence of refuges for sneakers in this species. FemaleC. microlepidotus deposited their entire clutch of about 9 eggs in one male territory. In contrast, femaleP. brieni divided their clutch of about 11 eggs among several males. After the final egg-release, femaleC. microlepidotus repeatedly approached their mate, with the mouth near the abdomen of the latter (nuzzling), but femaleP. brieni often departed without nuzzling. Males may eject sperm during nuzzling to fertilize eggs inside the female's mouth. However, maleP. brieni is also known to eject sperm near spawning females when the females are not nuzzling. Such behavior seems to be a male countermeasure against female mate infidelity, because males could not ensure paternity of eggs by ejecting sperm only during female nuzzling.     

Comparative Feeding Ecology of Two Sympatric Greenling Species, <Emphasis Type="Italic">Hexagrammos otakii</Emphasis> and <Emphasis Type="Italic">Hexagrammos agrammus</Emphasis> in Eelgrass <Emphasis Type="Italic">Zostera marina</Emphasis> Beds

Synopsis Feeding ecology was compared between sympatric greenling species, Hexagrammos otakii and H. agrammus in the eelgrass beds in Jindong Bay, Korea, from January to December. These two species had similar diets composed of crustaceans, polychaetes, gastropods and fishes; both species consumed primarily crustaceans throughout study periods. H. otakii, however, fed a greater proportion of polychaetes and fishes than H. agrammus. H. agrammus had a greater proportion of gastropods in their diets. The diet of both species underwent size-related changes; smaller individuals of H. otakii and H. agrammus consumed amphipods (gammarid amphipods and caprellid amphipods), while larger individuals of H. otakii ate polychaetes and fishes and those of H. agrammus fed mainly on gastropods and crabs. The diet of H. otakii underwent seasonal changes; H. otakii consumed mainly polychaetes and fishes during January and February 2002 but amphipods during March and May 2002. H. agrammus, however, ate mainly gastropods and crabs all seasons. H. otakii underwent also a significant diel changes that could be related to difference of prey availability. Thus the nocturnal emergence of gammarid amphipods, polychaetes and fishes explained their greater consumption by H. otakii. Dietary breadth of both species was lower in the smallest individuals (<5 cm SL) and in March and April 2002. This was due to the disproportionate dry mass attributable to the consumption of amphipods by both H. otakii and H. agrammus. Dietary overlap of both species was relatively moderate to high, in particular in <9.9 cm SL (0.62 – 0.71) from May to July 2002 (0.63 – 0.71). This is coincident with higher abundances of crabs, caridean shrimps and polychaetes in the study area, and it was assumed that these prey species were not limited resources. Higher dietary overlap was correlated with an abundance of a shared resource and did not indicate the interspecific competition between H. otakii and H. agrammus.     

Territorial male gobies respond aggressively to sneakers but do not adjust their sperm expenditure

We investigated whether mating behavior (sperm expenditure,courtship rate, and nest guarding) varied according to differentlevels of sperm competition in territorial males of two gobyspecies, the grass and the black gobies. We measured sperm expenditure(sperm released after 30 min from the beginning of the spawning),male courtship rate, and nest-guarding behavior in territorialmales of both species during simulated spawnings, in which wevaried the number of attending sneakers. Our results showedthat, in both species, territorial males adjusted their effortin nest guarding to the presence of rival sneakers by increasingthe time spent patrolling the territory and attacking the sneakers.In contrast, sperm expenditure and male courtship rate werenot influenced by the number of attending sneakers. These resultsare in agreement with those reported for other fish with alternativemating tactics and help to interpret previous inconsistenciesbetween theoretical predictions and measured levels of spermreleased at different levels of sperm competition by sneakersof the two gobiids studied here.     

Reproductive success in male sunbleak, a recent invasive fish species in the U.K.

The spawning behaviour of male nest guarding sunbleak Leucaspius delineatus , a recent invasive species in southern England, was studied and quantified for the first time. In the absence of physical differences between territorial and non‐territorial males ( i.e . colour, size, etc .), the reproductive behaviour of territorial males was analysed and related to reproductive success. The results showed that females preferred high‐courting and highly aggressive males. The initial cue in female mate choice, however, was based on courtship, while aggression was the decisive behavioural trait in influencing mate choice, providing a direct signal of physical condition and 'paternal competence'. Some males picked nest sites which were subsequently preferred by other males taking over the nest of a previous male ('communal nest'), with the new territorial male adopting the eggs already present at the nest. It appears that either due to female preference for nests already containing eggs or lower rates of sired egg predation by dilution among unrelated eggs, sunbleak males have adopted the mating strategy of allopaternal care.     

Comparative Study of Mitochondrial DNA in Two Greenling Species (Hexagrammidae: Pisces) and Their Hybrids from Peter the Great Bay (Sea of Japan)

Fragments of mtDNA from two greenling species, Hexagrammos octogrammusand H. agrammus, and their possible hybrids were amplified in PCR and examined using RFLP analysis. The inferred sequence difference between the species was 6.6%, which corresponds to about 3 Myr of their divergence. The reproductive contribution of males and females to the hybrids was estimated.     

Obstructive Role of the Dimorphic Sperm in a Non-copulatory Marine Sculpin, Hemilepidotus gilberti, to Prevent Other Males' Eusperm from Fertilization

In many species of animals, males normally produce parasperm (dimorphic sperm) along with eusperm (normal sperm) during spermatogenesis. In the present study, to clarify the role of parasperm of the non-copulatory sculpin Hemilepidotus gilberti, whose reproduction is characterized by polyandrous oviposition involving sneaking by neighboring territorial males, we observed the movements of parasperm. Parasperm could not move by themselves, but they were transported in solutions by passive movement due to collisions with actively swimming eusperm. In the viscous ovarian fluid (OF), which isolates eggs from seawater by covering them during spawning, parasperm did not exhibit any movement. However, they could be transported by eusperm movement in solutions with dissolved OF, partly because the viscosity of the fluid become lower. And then, in some solutions parasperm formed lumps. Lump formation of parasperm was also observed at the boundary surface of an egg mass where OF contacted seawater. Eusperm added experimentally to a solution in which parasperm were forming lumps were engulfed in the lumps and never escaped. Thus, lump formation of parasperm would be obstacles for the later arriving eusperm. Although lumps formed against both kin and non-kin eusperm, parasperm are thought to be available to overcome sperm competition which would occur during spawning that involves sneaking being almost concurrent with lump formation. The territorial male eusperm reach the eggs while his parasperm hinder other males' eusperm from reaching the eggs. Thus, we concluded that parasperm of H. gilberti play a role on protection of paternity by blocking rival eusperm physically from approaching eggs.     

The disruption of habitat isolation among three Hexagrammos species by artificial habitat alterations that create mosaic-habitat

In the coastal areas of Japan, three species of greenling (Hexagrammos spp.) can hybridize. In a natural reef setting we showed that Hexagrammos agrammus and H. octogrammus established their breeding territories in a shallow area where seaweed was abundant, whereas H. otakii established breeding territories in a deep area that was sparsely covered with seaweed. This difference in habitat use resulted in H. otakii being distributed separately from the other two species, thereby reducing the potential for hybridization. However, all the three species co-occurred in an artificial area near a breakwater. This area is characterized by steep slopes and complex stacked concrete structures, which create a mosaic-habitat consisting of a shallow environment with seaweed and a deep environment with sparse seaweed, allowing the three species to breed within a single area. Our results suggest that man-made structures can create an artificial mosaic-habitat that can disrupt habitat isolation and promote hybridization between species.     

Mate availability affects female choice in a fish with paternal care: female counterstrategies against male filial cannibalism

Theory predicts that individuals should adopt counterstrategies against intersexual conflict with their mating partners if the counterstrategies are effective and cost-efficient. In fishes, males with parental care often cannibalize their own offspring, which reduces the female’s fitness and creates intersexual conflicts. Males of the goby Rhinogobius flumineus cannibalize more eggs in the nest when they have access to additional females prior to spawning. Thus, it is predicted that females will strategically avoid spawning with males that have high mate availability. In the present study, we experimentally tested this prediction. When sexual pairs were placed in tanks, most females (control females; 21/22) successfully spawned inside the nest. In contrast, when a gravid female (stimulus female) that was housed in a small transparent cage was shown to the experiment pairs prior to spawning, only about half of the females (experiment females; 16/29) spawned inside the nest; the remaining females released unfertilized eggs outside of the nest. Moreover, experiment females infrequently accepted and followed males into nests, and delayed spawning more often than control females. R. flumineus females prefer males that court frequently. Indeed, experiment females that infrequently received courtship tended to spawn outside of the nest. However, infrequent courtship alone could not explain outside-nest spawning, delay in spawning, or the shorter stay of females in nests. These results imply that the presence of a stimulus female dampens female spawning with males. We suggest that R. flumineus females may strategically reject or hesitate to spawn with males that have high mate availability, and that this spawning avoidance may be a counterstrategy against male filial cannibalism.     

Effect of density on the reproductive behaviour of the territorial male demoiselle Chromis dispilus (Pisces: Pomacentridae)

Synopsis Characteristics of nest sites and reproductive behaviour of territorial male demoiselles Chromis dispilus were examined at sites of low, medium and high population density in northeastern New Zealand, by use of SCUBA diving and a remote underwater video system. Nest territories were closer together at high density (relative to areas of low population density) and this was accompanied by an increase in the frequency of spawning sequences and territorial defence against conspecifics. Territorial males were more aggressive during periods of display and spawning than during egg guarding. Fish from areas of low density spent more time on courtship display (signal jumps) and territorial defence against fish of other species, than demoiselles from areas of high population density. Territories and nest sizes tended to be largest in areas of medium population density. As in other damselfishes, population density is a major determinant of the frequency and intensity of reproductive behaviours.     

Interspecific territoriality in males of the tube blenny <Emphasis Type="Italic">Neoclinus bryope</Emphasis> (Actinopterygii: Chaenopsidae)

Territoriality was investigated in the tube blenny Neoclinus bryope (Actinopterygii: Chaenopsidae) at rocky intertidal areas of Banda Beach, Tateyama Bay, central Japan. Males used small holes as spawning nests, usually staying at the nest and maintaining the area while showing exclusive behaviors. Their home ranges were limited to areas within 30 cm distance from the nest for over 2 months. Four heterospecific fishes were threatened when they approached to within 6–14 cm of the nest holes, and two species of carnivorous snails were removed at points 0–30 cm from the nest entrance. There were no significant differences between the distances from the nest entrances to the points defended against fish and those used for foraging. As the four heterospecific species have similar feeding habits to those of N. bryope, the area defended against fishes may function as a foraging territory. At 24 h following the removal of nest owners, carnivorous snails had gathered to actively prey on eggs, indicating that the area defended against snails may function as a territory for protection against egg predators. Although the positions of females where males started courtship displays were significantly farther than the foraging points and the positions of threatened fishes, males displayed no territoriality against conspecific males. The fact that males did not leave the nest hole during the courtship suggests that it may be costly to maintain a courtship territory. These results show that males of tube blenny maintain territories for egg guarding and for protecting food resources around their nest holes in the spawning season.     

Black goby territorial males adjust their ejaculate's characteristics in response to the presence of sneakers

In many species, males can rapidly adjust their ejaculate performance in response to changing levels of sperm competition, an ability that is probably mediated by seminal fluid adaptive plasticity. In the black goby, Gobius niger, territorial males attach viscous ejaculate trails to the nest roof, from which sperm are slowly released into the water during the long-lasting spawning events. Sneaker males release their sperm in the vicinity of the nest, and territorial males try to keep them at a distance by patrolling their territory. We show here that territorial males'' ejaculate trails released a higher proportion of their sperm in the presence of a single sneaker, but this proportion decreased when there were three sneakers, an effect that is most likely mediated by a change in the seminal fluid composition. Field observations showed that when multiple sneaking attempts occurred, territorial males spent more time outside the nest, suggesting that ejaculation rate and territory defence are traded-off. Altogether, these results suggest that the adjustment of sperm release from the ejaculate may be strategic, guaranteeing a more continuous concentration of the territorial male''s sperm in the nest, although at a lower level, when he is engaged in prolonged territory defence outside the nest.     

Mate Choice and Mating Pattern in a Stream Goby of the Genus Rhinogobius

The mate choice and mating pattern of a benthic goby Rhinogobius sp. CB (cross band type) were investigated in the Kamo River, Shikoku, Japan. During the breeding season, gravid females assumed a nuptial color and either males or females initiated a courtship display. Males preferentially courted a female of similar size to lead her to his nest, whereas females courted more frequently when they encountered a large male. Eggs in any one nest were always at the same developmental stage. Sampling data of nesting males and females indicated that, in more than half the nests, males gathered more than one female before spawning. In some nests with eggs, two or three females had spent ovaries, indicating that the eggs were laid by multiple females within a short span of time. However, a comparison between the total number of eggs which females would spawn in one nest and the number of eggs actually deposited suggested that eggs were contributed by one female in most nests. This low level of polygyny in spite of multiple female availability is attributed to a limited available spawning area of the nest.     

Social organization and spawning in the Atlantic sharpnose puffer,Canthigaster rostrata (Tetraodontidae)

Synopsis Social organization and spawning in the sharpnose pufferCanthigaster rostrataere studied on a reef in the San Blas Islands, Panama. Sexes were dimorphic. In mixed coral and rubble habitat, females defended territories against other females and small males. From one to six female territories were included within the territories of certain large males. These haremic males visited their females and patrolled their territories throughout the day. Smaller, non-haremic males occupied territories or home ranges within or adjacent to those of haremic males or were wanderers. Spawning between a haremic male and a territorial female occurred within the female's territory. The female prepared an algal nest into which demersal eggs were deposited. There was no parental care. Eggs were spherical, translucent, and measured approximately 0.66 mm in diameter. Larvae were about 1.4 mm TL and closely resembled those of other species ofCanthigaster.     

Male Parasitic Spawning in Two Species of Triplefin Blenny (Tripterigiidae): Contrasts in Demography, Behaviour and Gonadal Characteristics

Axoclinus nigricaudus and A. carminalis are blennioid fishes from the Gulf of California, Mexico. Males show alternative mating tactics; territorial individuals attract females while non-territorial individuals parasitically spawn with territorial males' females. Demography and gonadal characteristics were related to the degree of parasitic spawning (sperm competition) within and between species. Males of both species showed bimodal size distributions that largely reflected the two tactics. Territorial males spawned much more frequently than non-territorial males, but parasitic spawning was significant. Non-territorial males were more common and rates of parasitic spawning were much higher in A. nigricaudus indicating that it experienced higher sperm competition. In agreement with sperm competition theory, relative testis weight was greater in A. nigricaudus. In A. nigricaudus, the majority of parasitic matings were by non-territorial males, whereas in A. carminalis, both male-types parasitised frequently. Accordingly, only in A. nigricaudus did non-territorial males have relatively heavier testis than territorial males. In both species the testicular gland, that produces accessory products for sperm transfer, was large in territorial males, but small or undeveloped in non-territorial males suggesting male-types differ in fertilisation mode.     

The courtship behaviour of the pipefish Nerophis lumbriciformis: reflections of an adaptation to intertidal life

The courtship behaviour of Nerophis lumbriciformis (Pisces: Syngnathidae) consists of three distinct phases (initial courtship, spawning, and embrace) marked by prominent behavioural changes. The first courtship phase is characterised by female quivering. Courtship activity increases from low to high levels of quivering, causing undulatory head movements in the female. In the second phase, the female transfers her eggs onto the male incubating ventral surface. During the final phase, the male wraps his body around the female (embrace). Females actively initiate courtship indicating that these pipefishes are courtship role reversed. The complete lack of swimming and vertical movements in the courtship behaviour of N. lumbriciformis, unlike in the behaviour of other syngnathid species, suggests an adaptation to intertidal conditions.     

Spawning of the sea ravenHemitripterus villosus in Peter the Great Bay, Sea of Japan

The spawning habits of the sea ravenHemitripterus villosus were investigated by scuba diving at shallow rocky bottom sites around Bolshoi Pelis Island (Peter the Great Bay, Sea of Japan). Spawning occurs in September, when the surface water temperature drops. The optimum spawning temperature is 17–18°C. The fish density in nest sites reaches 120/300 m². The mean individual fecundity is about 10000 eggs. Over 35% of the egg masses ofH. villosus are eaten by echinoderms, primarilyPatiria pectinifera andStrongylocentrotus nudus. Since fecundity is relatively low and parents do not take care of the egg masses, predation can strongly affect the abundance of this species.     

Feeding habits of the Alaska greenlingHexagrammos octogrammus in the coastal waters of amurskii bay (sea of Japan)

Feeding habits of the Alaska greenlingHexagrammos octogrammus were studied in the coastal waters of Amurskii Bay in April–October 1997–1998. The greenling is characterized by a wide feeding spectrum of no less then 56 fish and invertebrate species. Decapods predominate in the diets of all fish age groups. The feeding spectrum ofH. octogrammus broadens as the fish grows. From spring to autumn, the stomach filling index of first-year fish rises, while that of the other age groups decreases, reaching minimum values in the spawning period.     

Sound production during courtship and spawning of Oreochromis mossambicus: male–female and male–male interactions

The role of sound production of the Mozambique tilapia Oreochromis mossambicus in agonistic and mating interactions observed during hierarchy formation and in established groups was examined. Only territorial males produced sounds, during male–female and male–male courtship interactions and during pit-related activities ( e.g. dig, hover and still in the nest). Sound production rate was positively correlated with courting rate. Although sounds in other cichlids are typically emitted in early stages of courtship, O. mossambicus produced sounds in all phases, but especially during late stages of courtship, including spawning. It is suggested that the acoustic emissions in this species may play a role in advertising the presence and spawning readiness of males and in synchronizing gamete release.